Genital herpes is a recurrent, incurable sexually transmitted infection (STI) that is caused by infection with herpes simplex virus (HSV). After primary infection, HSV establishes latency in the sensory neurons that innervate the genital tract. Periodic reactivation of the virus within the peripheral nervous system (PNS) and subsequent replication in the genital epithelium causes the recurrent lesions associated with the disease. Thus, protection of the PNS is a key factor in preventing the disease caused by HSV. Recently, our lab developed a novel vaccine strategy called prime and pull, which establishes a pool of tissue-resident memory CD8 T cells (CD8 TRM) in the female genital tract that provides immunity against genital herpes by protecting sensory neurons from HSV infection. The overall goal of this proposal will examine the novel mechanisms by which CD8 TRM potentiate antiviral mechanisms that protect the PNS from primary infection with HSV. Currently, we are using the prime and pull system to begin determining the components and kinetics of the immune response that lead to neuroprotection against HSV infection in the genital tract. Once the independent phase of my research begins, I will continue to use prime and pull to dissect the cellular and molecular requirements for CD8 TRM-mediated neuroprotective responses as described in this K22 research proposal. Aim 1 will elucidate the activation and functional requirements for CD8 TRM-mediated neuroprotective responses. An in vitro co-culture system will also be developed to investigate the hypothesis that CD8 TRM directly engage neuron-intrinsic antiviral pathways to restrict HSV infection. This co-culture system will be used in Aim to dissect the neuron-extrinsic and -intrinsic antiviral pathways that are mediated by CD8 TRM. While this K22 proposal advances my longstanding interest in the antiviral role of memory CD8 T cells, it will also begin to shift my focus to the interface between immune and nervous system. Dr. Akiko Iwasaki, my current mentor, will provide scientific guidance as well as advice on transitioning to an independent position. Along with Dr. Iwasaki, an advisory committee will oversee my scientific progress and guide my career development. The in vitro culture system will be developed in close collaboration with Dr. Lynn Enquist, who is a leading expert in neurovirulent viruses. Overall, the long- term goal of my future research, as established by this K22 proposal, will be to increase understanding of how tissue-resident immune cells and the peripheral nervous system interact to mediate protection against neurotropic pathogens.